The biopharmaceutical industry has shown significant interest in developing continuous manufacturing processes for therapeutic protein production. This is partly due to advantages compared to fed-batch production methods, which include higher productivity, increased product quality, reduction in bioreactor sizes, and potentially better utilization of production infrastructure. Continuous manufacturing approaches have clear advantages for processes that may be difficult to scale up without inducing changes to the therapeutic product. Despite this progress, continuous manufacturing approaches have yet to be applied to viral vector manufacturing. The growing cell and gene therapy industry has three key needs that may be addressed by continuous manufacturing of viral vectors. First, the anticipated worldwide demand for viral vectors cannot feasibly be met using current batch production infrastructure, necessitating the use of new and alternative manufacturing approaches. Second, continuous manufacturing could potentially allow rapid production of quantities of vectors which would enable more rapid initiation of clinical trials, thus increasing speed to market for novel gene therapy products. Third, as biologic products become larger and more complex (with cell therapies being more complex than viral therapies being more complex than proteins), scaling up from clinical scale to commercial scale manufacturing processes becomes challenging. Manufacturing changes due to scale up (e.g. increasing from 500L to 2000L and the accompanying changes in agitation rate) can lead to unanticipated changes in the product quality and final clinical performance. Therefore, developing a large-scale process that produces a comparable product as well as proving that comparability is both time consuming and resource intensive. To help address these challenges, MIT is proposing the development and demonstration of a continuous upstream viral vector manufacturing platform. This will be accomplished through three distinct aims: first, we will develop a first principles mathematical model for continuous viral vector cell culture unit operation process design; second, we will demonstrate the application of novel analytics for the in-line measurement of plasmid transfection and vector production parameters; and third, we will leverage these learnings to demonstrate the continuous cell culture production of viral vectors. At the end of the project we will have exhibited a generic approach for continuous viral vector manufacturing that can be applied to other viral vectors of interest for the production of either gene or gene-modified cell therapies.